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1.1. INTRODUCTION The objective of many bioassay methods is to selectively quantitate a single biomolecule, such as a particular enzyme or antibody, or to determine the presence or absence of a known DNA sequence in an unknown sample. Methods for these very selective assays will be considered in later chapters. When faced with a true unknown, for example, during the isolation or puri cation of a biomolecule, it can be important to characterize the unknown matrix, or the components in the unknown solution that are present along with the species of interest. This involves the estimation of the total quantity of the different types of biomolecules. Biochemists often estimate the total quantity of protein and nucleic acid in an unknown by the nomograph method.1 In this method, the absorbance of the unknown solution in a 1-cm cuvette is measured at 260 and 280 nm. The nomograph (Fig. 1.1) is then used to estimate concentrations. The nomograph method is rapid and involves no chemical derivatization, but suffers from several disadvantages. Interferences result from any species present in the unknown, other than protein or nucleic acid, that absorb at these wavelengths. Phenol, for example, which is used at high concentrations during the puri cation of nucleic acid, absorbs at both 260 and 280 nm and leads to overestimation of total nucleic acid. Furthermore, the nomograph method does not distinguish between deoxyribonucleic acid (DNA) and ribonucleic acid (RNA), and is useful only for samples containing relatively high concentrations of protein and nucleic acid. In this chapter, we will consider simple colorimetric and uorometric methods for the quantitation of total protein, DNA, RNA, carbohydrate, and free fatty acid. All of the methods are based on the generation of a chromophore or uorophore by a selective chemical reaction.

325 Supervising exchanges 33 Automating business processes 331 Modeling business processes 332 Executing business processes 333 Supervising business processes 34 Business process and integration: mediation and exchange 341 Business process level and integration level 342 Mediation process sub-level 343 Exchange process sub-level 344 Interaction between the sub-levels 345 Interaction between integration and business process (BPM) 35 Choosing the exchange architecture 351 Synchronous/asynchronous communication 352 Architecture: centralized or distributed 41 Integrating a single application 411 Exchange cartography 412 The integration platform 42 IT infrastructure projects 421 Urbanization of information systems 422 IT exchange infrastructure 43 Integrating inter-enterprise exchanges 431 Exchanging electronic documents (EDI) 432 XML standards 433.

Contents Failure Modes Hardware Environmental and Physical Failures Network Failures File and Print Server Failures Database System Failures Web and Application Server Failures Denial-of-Service Attacks Confidence in Your Measurements Renewability Sigmas and Nines Key Points 3 The Value of Availability What Is High Availability The Costs of Downtime Direct Costs of Downtime Indirect Costs of Downtime The Value of Availability Example 1: Clustering Two Nodes Example 2: Unknown Cost of Downtime The Availability Continuum The Availability Index The Lifecycle of an Outage Downtime Lost Data Degraded Mode Scheduled Downtime Key Points The Politics of Availability Beginning the Persuasion Process Start Inside Then Go Outside Legal Liability Cost of Downtime Start Building the Case Find Allies Which Resources Are Vulnerable Develop a Set of Recommendations Your Audience Obtaining an Audience Know Your Audience Delivering the Message The Slide Presentation The Report After the Message Is Delivered Key Points 20 20 21 23 24 24 26 27 28 28 29 30 31 31 34 34 36 37 42 46 47 51 52 53 55 57 57 60 61 61 62 63 63 64 65 65 66 68 69 69 70 70 70 71 73 73

Bianalytical Chemistry, by Susan R. Mikkelsen and Eduardo Corton ISBN 0-471-54447-7 Copyright # 2004 John Wiley & Sons, Inc.

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